Low storage temperature affects quality and volatile compounds in fresh tomatoes.

To investigate the impact of low temperature on the quality and flavor of ripe red tomatoes, we analyzed transcriptomes and volatile metabolomes of ripe red fruits stored at 0 °C and 20 °C for 8 days. The results showed that 0 °C maintained the sugar content by increasing the expression of sucrose synthetase (SUS) and sucrose transporter (SUT). Low expression of aroma synthesis-related genes, such as alcohol dehydrogenase 1 (ADH1), amino acid decarboxylase 1 A (AADC1A), and branched-chain amino acid aminotransferase 2 (BCAT2), were associated with reduced levels of pentanal, hexanal, 3-methylbutanal, 2-methylbutanal, and 2-phenylethanol. Additionally, the expression of pectinesterase (PE), beta-galactosidase (ß-GAL), and beta-glucosidase (ß-Glu), as well as phytoene synthase1 (PSY1) involved in carotenoid synthesis, was inhibited, thereby maintaining fruits texture and color. Furthermore, storage at 0 °C induced the expression of numerous genes regulating antioxidant and heat shock proteins, which further preserved the postharvest quality of tomatoes.

Transcriptomic, metabolomic, and ATAC-seq analysis reveal the regulatory mechanism of senescence of post-harvest tomato fruit.

Several physiological changes occur during fruit storage, which include the regulation of genes, metabolisms and transcription factors. In this study, we compared 'JF308' (a normal tomato cultivar) and 'YS006' (a storable tomato cultivar) to determine the difference in accumulated metabolites, gene expression, and accessible chromatin regions through metabolome, transcriptome, and ATAC-seq analysis. A total of 1006 metabolites were identified in two cultivars. During storage time, sugars, alcohols and flavonoids were found to be more abundant in 'YS006' compared to 'JF308' on day 7, 14, and 21, respectively. Differentially expressed genes, which involved in starch and sucrose biosynthesis were observed higher in 'YS006'. 'YS006' had lower expression levels of CesA (cellulose synthase), PL (pectate lyase), EXPA (expansin) and XTH (xyglucan endoglutransglucosylase/hydrolase) than 'JF308'. The results showed that phenylpropanoid pathway, carbohydrate metabolism and cell wall metabolism play important roles in prolonging the shelf life of tomato (Solanum lycopersicum) fruit. The ATAC-seq analysis revealed that the most significantly up-regulated transcription factors during storage were TCP 2,3,4,5, and 24 in 'YS006' compared to 'JF308' on day 21. This information on the molecular regulatory mechanisms and metabolic pathways of post-harvest quality changes in tomato fruit provides a theoretical foundation for slowing post-harvest decay and loss, and has theoretical importance and application value in breeding for longer shelf life cultivars.

Microbial antagonists to biologically control postharvest decay and preserve fruit quality.

Postharvest waste due to decay of fruits and vegetables negatively affects food security, while at the same time control of decay and therefore waste can be limited because of consumer concerns about use of synthetic chemicals. Use of antagonistic microorganisms is an eco-friendly technique that represents a promising alternative approach to the use of chemical methods. Understanding the interactions between antagonists and the fruit microbiome will enable the discovery of new methods to reduce postharvest waste. This article reviews different microbial agents, fungi, bacteria and yeasts that could control decay. Recent developments in the use of microorganisms for preserving postharvest fruit quality, formulation of effective antagonists, and the commercialization steps are also discussed. Antagonists control decay through either direct or indirect mechanisms while preserving the appearance, flavor, texture and nutritional value of horticultural products. Microorganisms do not fully control pathogens, and therefore they are usually used with other treatments or have their biocontrol ability modified through genetic manipulations. Despite of these limitations, commercialization of biocontrol products based on antagonists with required stability and biocontrol potential is occurring. Biocontrol of postharvest decay and waste agent is promising technology for fruit and vegetable industries. Further study is necessary to better understand mechanisms and increasing efficiency of this method.

Revealing the Specific Regulations of Brassinolide on Tomato Fruit Chilling Injury by Integrated Multi-Omics.

Tomato fruit is susceptible to chilling injury (CI) when stored at low temperatures, limiting its storage potential, and resulting in economic loss if inappropriate temperatures are used. Brassinolide (BR) is a plant growth regulator that is known to decrease the susceptibility of fruit to CI. In this study, transcriptome, metabolome, and proteome analysis revealed the regulation mechanism of BR treatment in alleviating tomato fruit CI. The results showed that the differentially expressed metabolites mainly included amino acids, organic acids, carbohydrates, and lipids. Differentially expressed genes (DEGs) were involved in plant cold stress response (HSFA3, SHSP, and TPR), fruit redox process (POD, PAL, and LOX), related to the fruit texture (CESA, ß-Gal, and PAE), plant hormone signal transduction (ACS3, ARF, and ERF,), transcription factors (TCP, bHLH, GATA). Moreover, differentially expressed proteins were associated with fruit texture (CESA, PE, PL, and CHI), plant oxidation processes (LOX, GPX, CAT, and POD), plant cold stress response (HSF, HSP20, HSP70, and HSP90B), plant hormone signal transduction (BSK1 and JAR1) and transcription factors (WRKY and MYB). Our study showed that BR alleviates CI symptoms of tomato fruit by regulating LOX in the α-linolenic acid metabolism pathway, enhancing jasmonic acid-CoA (JA-CoA) synthesis, inhibiting cell wall and membrane lipid damage. The results provided a theoretical basis for further study on the CI mechanism of tomato fruit.

The leaf senescence-promoting transcription factor AtNAP activates its direct target gene CYTOKININ OXIDASE 3 to facilitate senescence processes by degrading cytokinins.

Cytokinins (CKs) are a class of adenine-derived plant hormones that plays pervasive roles in plant growth and development including cell division, morphogenesis, lateral bud outgrowth, leaf expansion and senescence. CKs as a "fountain of youth" prolongs leaf longevity by inhibiting leaf senescence, and therefore must be catabolized for senescence to occur. AtNAP, a senescence-specific transcription factor has a key role in promoting leaf senescence. The role of AtNAP in regulating CK catabolism is unknown. Here we report the identification and characterization of AtNAP-AtCKX3 (cytokinin oxidase 3) module by which CKs are catabolized during leaf senescence in Arabidopsis. Like AtNAP, AtCKX3 is highly upregulated during leaf senescence. When AtNAP is chemically induced AtCKX3 is co-induced; and when AtNAP is knocked out, the expression of AtCKX3 is abolished. AtNAP physically binds to the cis element of the AtCKX3 promoter to direct its expression as revealed by yeast one-hybrid assays and in planta experiments. Leaves of the atckx3 knockout lines have higher CK concentrations and a delayed senescence phenotype compared with those of WT. In contrast, leaves with inducible expression of AtCKX3 have lower CK concentrations and exhibit a precocious senescence phenotype compared with WT. This research reveals that AtNAP transcription factor-AtCKX3 module regulates leaf senescence by connecting two antagonist plant hormones abscisic acid and CKs.

Meta-analysis of the effects of 1-methylcyclopropene (1-MCP) treatment on climacteric fruit ripening.

1-Methylcyclopropene (1-MCP) is an inhibitor of ethylene perception that is widely used to maintain the quality of several climacteric fruits during storage. A large body of literature now exists on the effects of 1-MCP on climacteric fruit ripening for different species and environmental conditions, presenting an opportunity to use meta-analysis to systematically dissect these effects. We classified 44 ripening indicators of climacteric fruits into five categories: physiology and biochemistry, quality, enzyme activity, color, and volatiles. Meta-analysis showed that 1-MCP treatment reduced 20 of the 44 indicators by a minimum of 22% and increased 6 indicators by at least 20%. These effects were associated with positive effects on delaying ripening and maintaining quality. Of the seven moderating variables, species, 1-MCP concentration, storage temperature and time had substantial impacts on the responses of fruit to 1-MCP treatment. Fruits from different species varied in their responses to 1-MCP, with the most pronounced responses observed in rosaceous fruits, especially apple, European pear fruits, and tropical fruits. The effect of gaseous 1-MCP was optimal at 1 µl/l, with a treatment time of 12-24 h, when the storage temperature was 0 °C for temperate fruits or 20 °C for tropical fruits, and when the shelf temperature was 20 °C, reflecting the majority of experimental approaches. These findings will help improve the efficacy of 1-MCP application during the storage of climacteric fruits, reduce fruit quality losses and increase commercial value.

Primary Metabolism in Fresh Fruits During Storage.

The extension of commercial life and the reduction of postharvest losses of perishable fruits is mainly based on storage at low temperatures alone or in combination with modified atmospheres (MAs) and controlled atmospheres (CAs), directed primarily at reducing their overall metabolism thus delaying ripening and senescence. Fruits react to postharvest conditions with desirable changes if appropriate protocols are applied, but otherwise can develop negative and unacceptable traits due to the onset of physiological disorders. Extended cold storage periods and/or inappropriate temperatures can result in development of chilling injuries (CIs). The etiology, incidence, and severity of such symptoms vary even within cultivars of the same species, indicating the genotype significance. Carbohydrates and amino acids have protective/regulating roles in CI development. MA/CA storage protocols involve storage under hypoxic conditions and high carbon dioxide concentrations that can maximize quality over extended storage periods but are also affected by the cultivar, exposure time, and storage temperatures. Pyruvate metabolism is highly reactive to changes in oxygen concentration and is greatly affected by the shift from aerobic to anaerobic metabolism. Ethylene-induced changes in fruits can also have deleterious effects under cold storage and MA/CA conditions, affecting susceptibility to chilling and carbon dioxide injuries. The availability of the inhibitor of ethylene perception 1-methylcyclopropene (1-MCP) has not only resulted in development of a new technology but has also been used to increase understanding of the role of ethylene in ripening of both non-climacteric and climacteric fruits. Temperature, MA/CA, and 1-MCP alter fruit physiology and biochemistry, resulting in compositional changes in carbon- and nitrogen-related metabolisms and compounds. Successful application of these storage technologies to fruits must consider their effects on the metabolism of carbohydrates, organic acids, amino acids and lipids.

Characterization of Fruit Quality Attributes and Cell Wall Metabolism in 1-Methylcyclopropene (1-MCP)-Treated 'Summer King' and 'Green Ball' Apples During Cold Storage.

This study aimed to elucidate whether 1-methylcyclopropene (1-MCP) treatment delays the fruit softening mechanism associated with the fruit quality of the newly released apple cultivars "Summer King" and "Green Ball" during cold storage. For both cultivars, the fruit treated with 1-MCP exhibited lower internal ethylene concentration, higher firmness, and higher titratable acidity relative to the control fruit, in association with less fruit softening. In addition, the treated fruit significantly delayed fresh weight loss and reduction of soluble solids content, especially in "Green Ball." Moreover, slower degradation of cell wall components (water-soluble pectin, sodium carbonate-soluble pectin, hemicellulose, and cellulose) was also observed in the treated fruit in comparison to the control fruit. Similarly, the enzymatic activities (of polygalacturonase, pectin methylesterase, cellulase, ß-galactosidase, and α-L-arabinofuranosidase) that cause cell wall degradation were relatively lower in the treated fruit than in the control fruit for both cultivars, which was further proved by transcriptional analysis of the genes encoding the enzymes. Overall, the results suggested that the usage of 1-MCP is useful to delay fruit softening of the two cultivars during cold storage by delaying the degradation of cell wall components and enzymatic activities of cell wall hydrolysis.

Delayed response to cold stress is characterized by successive metabolic shifts culminating in apple fruit peel necrosis.

BACKGROUND: Superficial scald is a physiological disorder of apple fruit characterized by sunken, necrotic lesions appearing after prolonged cold storage, although initial injury occurs much earlier in the storage period. To determine the degree to which the transition to cell death is an active process and specific metabolism involved, untargeted metabolic and transcriptomic profiling was used to follow metabolism of peel tissue over 180 d of cold storage. RESULTS: The metabolome and transcriptome of peel destined to develop scald began to diverge from peel where scald was controlled using antioxidant (diphenylamine; DPA) or rendered insensitive to ethylene using 1-methylcyclopropene (1-MCP) beginning between 30 and 60 days of storage. Overall metabolic and transcriptomic shifts, representing multiple pathways and processes, occurred alongside α-farnesene oxidation and, later, methanol production alongside symptom development. CONCLUSIONS: Results indicate this form of peel necrosis is a product of an active metabolic transition involving multiple pathways triggered by chilling temperatures at cold storage inception rather than physical injury. Among multiple other pathways, enhanced methanol and methyl ester levels alongside upregulated pectin methylesterases are unique to peel that is developing scald symptoms similar to injury resulting from mechanical stress and herbivory in other plants.

Gene expression and metabolism preceding soft scald, a chilling injury of 'Honeycrisp' apple fruit.

BACKGROUND: 'Honeycrisp' is an apple cultivar that is susceptible to soft scald, a chilling injury expressed as necrotic patches on the peel. Improved understanding of metabolism associated with the disorder would improve our understanding of soft scald and contribute to developing more effective management strategies for apple storage. It was expected that specific gene expression and specific metabolite levels in the peel would be linked with soft scald risk at harvest and/or specific time points during cold storage. RESULTS: Fruit from nine 'Honeycrisp' apple orchards that would eventually develop different incidences of soft scald between 4 and 8 weeks of cold air storage were used to contrast and determine differential transcriptomic and metabolomic changes during storage. Untargeted metabolic profiling revealed changes in a number of distinct pathways preceding and concurrent with soft scald symptom development, including elevated γ-aminobutryic acid (GABA), 1-hexanol, acylated steryl glycosides, and free p-coumaryl acyl esters. At harvest, levels of sesquiterpenoid and triterpenoid acyl esters were relatively higher in peel of fruit that did not later develop the disorder. RNA-seq driven gene expression profiling highlighted possible involvement of genes and associated metabolic processes with soft scald development. These included elevated expression of genes involved in lipid peroxidation and phenolic metabolism in fruit with soft scald, and isoprenoid/brassinosteroid metabolism in fruit that did not develop soft scald. Expression of other stress-related genes in fruit that developed soft scald included chlorophyll catabolism, cell wall loosening, and lipid transport while superoxide dismutases were up-regulated in fruit that did not develop the disorder. CONCLUSIONS: This study delineates the sequential transcriptomic and metabolomic changes preceding soft scald symptom development. Changes were differential depending on susceptibility of fruit to the disorder and could be attributed to key stress related and mediating pathways.

Variations in zonal fruit starch concentrations of apples - a developmental phenomenon or an indication of ripening?

Patterns of starch hydrolysis in stem, equatorial, and calyx zones of 'Honeycrisp' and 'Empire' apples (Malus sylvestris (L.) Mill var. domestica (Borkh.) Mansf.) during maturation and ripening, and in 'Gala' apples in response to propylene or 1-methylcyclopropene (1-MCP) treatments after harvest, were studied. Differences in zonal starch concentrations were found for 'Empire' and 'Gala' fruits, but not for 'Honeycrisp'. During maturation and ripening of 'Empire', the concentration of starch was highest in the calyx end and lowest in the stem region. Differences in rates of starch hydrolysis among zones were not detected. 'Honeycrisp' and 'Empire' had the highest concentration of sorbitol in the calyx region, whereas it was highest in the stem-end region in 'Gala'. The distribution differences of glucose, fructose, and sucrose were similar in all three cultivars; higher fructose and glucose concentrations in the stem region, and higher sucrose concentrations in the calyx end of the fruit. Postharvest treatment of 'Gala' with propylene did not affect the internal ethylene concentration of the fruit but 1-MCP markedly inhibited it. Starch concentrations were highest in the calyx end but gradients of starch among zones were not changed by postharvest treatment. The rate of hydrolysis was slowed by 1-MCP treatment, but was unaffected by propylene. Postharvest treatments influenced sorbitol, glucose, and fructose concentrations. Patterns of starch concentration among the zones did not confirm differences in ripening, but reflected its uneven distribution throughout the fruit during development. Therefore, measured differences in zonal starch are most likely related to starch accumulation during fruit development, rather than differences in rates of starch degradation during ripening.

Chilling-related cell damage of apple (Malus × domestica Borkh.) fruit cortical tissue impacts antioxidant, lipid and phenolic metabolism.

'Soggy breakdown' (SB) is an internal flesh disorder of 'Honeycrisp' apple (Malus × domestica Borkh.) fruit that occurs during low temperature storage. The disorder is a chilling injury (CI) in which visible symptoms typically appear after several weeks of storage, but information about the underlying metabolism associated with its induction and development is lacking. The metabolic profile of flesh tissue from wholly healthy fruit and brown and healthy tissues from fruit with SB was characterized using gas chromatography-mass spectrometry (GC-MS) and liquid chromatograph-mass spectrometry (LC-MS). Partial least squares discriminant analysis (PLS-DA) and correlation networks revealed correlation among ester volatile compounds by composition and differences in phytosterol, phenolic and putative triacylglycerides (TAGs) metabolism among the tissues. anova-simultaneous component analysis (ASCA) was used to test the significance of metabolic changes linked with tissue health status. ASCA-significant components included antioxidant compounds, TAGs, and phytosterol conjugates. Relative to entirely healthy tissues, elevated metabolite levels in symptomatic tissue included γ-amino butyric acid, glycerol, sitosteryl (6'-O-palmitoyl) ß-d-glucoside and sitosteryl (6'-O-stearate) ß-d-glucoside, and TAGs containing combinations of 16:0, 18:3, 18:2 and 18:1 fatty acids. Reduced metabolite levels in SB tissue included 5-caffeoyl quinate, ß-carotene, catechin, epicatechin, α-tocopherol, violaxanthin and sitosteryl ß-d glucoside. Pathway analysis indicated aspects of primary metabolism differed according to tissue condition, although differences in metabolites involved were more subtle than those of some secondary metabolites. The results implicate oxidative stress and membrane disruption processes in SB development and constitute a diagnostic metabolic profile for the disorder.

Transcriptomic events associated with internal browning of apple during postharvest storage.

BACKGROUND: Postharvest ripening of apple (Malus x domestica) can be slowed down by low temperatures, and a combination of low O2 and high CO2 levels. While this maintains the quality of most fruit, occasionally storage disorders such as flesh browning can occur. This study aimed to explore changes in the apple transcriptome associated with a flesh browning disorder related to controlled atmosphere storage using RNA-sequencing techniques. Samples from a browning-susceptible cultivar ('Braeburn') were stored for four months under controlled atmosphere. Based on a visual browning index, the inner and outer cortex of the stored apples was classified as healthy or affected tissue. RESULTS: Over 600 million short single-end reads were mapped onto the Malus consensus coding sequence set, and differences in the expression profiles between healthy and affected tissues were assessed to identify candidate genes associated with internal browning in a tissue-specific manner. Genes involved in lipid metabolism, secondary metabolism, and cell wall modifications were highly modified in the affected inner cortex, while energy-related and stress-related genes were mostly altered in the outer cortex. The expression levels of several of them were confirmed using qRT-PCR. Additionally, a set of novel browning-specific differentially expressed genes, including pyruvate dehydrogenase and 1-aminocyclopropane-1-carboxylate oxidase, was validated in apples stored for various periods at different controlled atmosphere conditions, giving rise to potential biomarkers associated with high risk of browning development. CONCLUSIONS: The gene expression data presented in this study will help elucidate the molecular mechanism of browning development in apples at controlled atmosphere storage. A conceptual model, including energy-related (linked to the tricarboxylic acid cycle and the electron transport chain) and lipid-related genes (related to membrane alterations, and fatty acid oxidation), for browning development in apple is proposed, which may be relevant for future studies towards improving the postharvest life of apple.

Understanding development and ripening of fruit crops in an 'omics' era.

Next generation sequencing has revolutionized plant biology. Not only has our understanding of plant metabolism advanced using model systems and modern chromatography, but application of 'omics'-based technology has been widely extended to non-model systems as costs have plummeted and efficiency increased. As a result, important fundamental questions relating to important horticultural crops are being answered, and novel approaches with application to industry are in progress. Here we review recent research advances on development and ripening of fruit crops, how next generation sequencing approaches are driving this advance and the emerging future landscape.

Arabidopsis AtNAP regulates fruit senescence.

Arabidopsis has been used as a model system to study many aspects of plant growth and development. However, fruit senescence in Arabidopsis has been less investigated and the underlying molecular and hormonal (especially ethylene) regulatory mechanisms are not well understood. It is reported here that the Arabidopsis silique has characteristics of a climacteric fruit, and that AtNAP, a NAC family transcription factor gene whose expression is increased with the progression of silique senescence, plays an important role in its senescence. Silique senescence was delayed for 4-5 d in the atnap knockout mutant plants. The ethylene climacteric was delayed for 2 d in the atnap silique and the associated respiratory climacteric was suppressed. Exogenous ethylene stimulated respiration in the wild type, but not in the atnap mutant. The decoupling of the ethylene and respiratory climacterics in the atnap mutant suggests that AtNAP is required for ethylene stimulation of respiration. qPCR analyses revealed that the expression patterns of genes involved in ethylene biosynthesis, perception, and signalling, ACS2, ETR1, CTR1, EIN2, EIN3, and ERF1, were also altered in the atnap mutant. The effects of exogenous ABA, SA, 6-BA, and NAA on ethylene production and respiration in siliques of the wild type and atnap mutant were also investigated. A model involving ABA-AtNAP-controlled stomatal opening in regulating ethylene-stimulated respiration in fruit senescence is presented.

Fermentation and malate metabolism in response to elevated CO2 concentrations in two strawberry cultivars.

Concentrations of acetaldehyde, ethanol, ethyl acetate (EA), organic acids and activities and gene expression of alcohol dehydrogenase (ADH; EC 1.1.1.1), pyruvate decarboxylase (PDC; EC 4.1.1.1), alcohol acyltransferase (AAT; EC 1.4.1.14), malate dehydrogenase (MDH; EC 1.1.1.37), malic enzyme (ME; EC 1.1.1.40) and glutamate dehydrogenase (EC 1.4.1.14) were investigated in two strawberry (Fragaria x ananassa Duch) cultivars with different responses to CO(2) during storage. 'Jewel' fruit treated with CO(2) accumulated acetaldehyde and ethanol but little EA, while 'Cavendish' accumulated little acetaldehyde or ethanol but accumulated EA. In CO(2)-treated fruit, PDC activity was positively correlated with EA accumulation in 'Jewel' but not in 'Cavendish', while no differential effect of atmosphere was observed on its gene expression. ADH activity and gene expression show a correlation with ethanol accumulation in 'Cavendish'. In 'Jewel', there was a positive correlation between ADH gene expression and enzyme activity; however, this correlation does not explain ethanol accumulation in this cultivar. EA accumulation did not show any correlation with AAT activity and gene expression in any of the cultivars. Succinate concentrations were highest and those of malate lowest in CO(2)-treated fruit of both cultivars, but MDH and ME activities were not affected by CO(2). Gene expression of MDH and ME were not affected by atmosphere in 'Cavendish', although in 'Jewel' the MDH expression was slightly lower in CO(2)- than air-treated fruit. The results of this study show that differences in fermentation products and malate accumulation in CO(2)-treated strawberry fruit are not consistently correlated with enzyme activities and gene expression.

Inhibition of the ethylene response by 1-MCP in tomato suggests that polyamines are not involved in delaying ripening, but may moderate the rate of ripening or over-ripening.

Ethylene initiates the ripening and senescence of climacteric fruit, whereas polyamines have been considered as senescence inhibitors. Ethylene and polyamine biosynthetic pathways share S-adenosylmethionine as a common intermediate. The effects of 1-methylcyclopropene (1-MCP), an inhibitor of ethylene perception, on ethylene and polyamine metabolism and associated gene expression was investigated during ripening of the model climacteric fruit, tomato (Solanum lycopersicum L.), to determine whether its effect could be via polyamines as well as through a direct effect on ethylene. 1-MCP delayed ripening for 8 d compared with control fruit, similarly delaying ethylene production and the expression of 1-aminocyclopropane-1-carboxylic acid (ACC)-synthase and some ethylene receptor genes, but not that of ACC oxidase. The expression of ethylene receptor genes returned as ripening was reinitiated. Free putrescine contents remained low while ripening was inhibited by 1-MCP, but increased when the fruit started to ripen; bound putrescine contents were lower. The activity of the putrescine biosynthetic enzyme, arginine decarboxylase, was higher in 1-MCP-treated fruit. Activity of S-adenosylmethionine-decarboxylase peaked at the same time as putrescine levels in control and treated fruit. Gene expression for arginine decarboxylase peaked early in non-treated fruit and coincident with the delayed peak in putrescine in treated fruit. A coincident peak in the gene expression for arginase, S-adenosylmethionine-decarboxylase, and spermidine and spermine synthases was also seen in treated fruit. No effect of treatment on ornithine decarboxylase activity was detected. Polyamines are thus not directly associated with a delay in tomato fruit ripening, but may prolong the fully-ripe stage before the fruit tissues undergo senescence.

Isolation and characterization of a lipid transfer protein expressed in ripening fruit of Capsicum chinense.

A novel LTP (CcLTP) from a Capsicum chinense cv Habanero was isolated from a fruit-specific SSH library. While this gene shares similarity with other LTPs, it is considerably larger than any lipid transfer protein reported to date and has a neutral predicted pI. CcLTP is consistently expressed in seedlings from three Capsicum species. It is also present at very high levels in ripening and mature fruit in C. chinense, but not in fruit of any C. annuum or C. frutescens varieties examined. We have obtained 3.8 kb of sequence containing the CcLTP gene and isolated two forms of mRNA transcripts which result from an alternative splicing event. Both transcripts are full-length cDNAs with putative open reading frames of 492 bp and 519 bp, encoding proteins of 164 and 173 amino acids, respectively, which differ only by an insertion of 9 amino acids. Both splice variants are detected consistently via RT-PCR. A 19 bp deletion in the promoter region differentiates C. chinense CcLTP from that of C. annuum and C. frutescens. The protein and its expression are characterized in C. chinense fruit, and a possible role in pepper fruit ripening and maturation is discussed.

1-Methylcyclopropene interactions with diphenylamine on diphenylamine degradation, alpha-farnesene and conjugated trienol concentrations, and polyphenol oxidase and peroxidase activities in apple fruit.

1-Methylcyclopropene (1-MCP) is a new technology that is applied commercially to inhibit ethylene action in apple fruit, but its interactions with existing technologies such as diphenylamine (DPA) for control of superficial scald development in fruit during and after storage is unknown. To investigate possible interactions between 1-MCP and DPA, Delicious apples were untreated or treated with 2 g L(-1) DPA, and then with or without 1 microL L(-1) 1-MCP. Ethylene production and respiration rates of fruit were measured immediately following treatment, and fruit was stored at 0.5 degrees C for 12 weeks. Internal ethylene concentrations (IEC), alpha-farnesene and conjugated trienol (CTol) concentrations, activities of peroxidase and polyphenol oxidase (PPO), and DPA levels in the skin of the fruit were measured at intervals during storage. 1-MCP reduced the rate of DPA loss from peel tissue so that by 12 weeks of storage concentrations of the chemical were 25% higher than in untreated fruit. 1-MCP, with and without DPA, markedly inhibited ethylene production and respiration rates, maintained low IEC and alpha-farnesene and CTol concentrations, while DPA had little effect on these factors except inhibition of CTol accumulation. Treatment effects on peroxidase and PPO activities were inconsistent.